Serum biochemistry profile in newborn Senepol and crossbred Holstein x Gir calves aged three to five days in Uberlândia, Minas Gerais, Brazil / Perfil bioquímico sérico de bezerros neonatos da raça Senepol e mestiços Holandês x Gir de três a cinco dias de idade no município de Uberlândia, MG, Brasil

This study aimed to evaluate the physiological variations in the serum biochemical profile of neonate (3-5 days old) Senepol and crossbreed Holstein x Gir calves in Uberlândia, state of Minas Gerais, Brazil, and to compare values between breeds and sexes. In total, 168 healthy Senepol (n = 80, 28 females and 52 males) and crossbred Holstein x Gir (n = 88, 49 females and 39 males) calves were evaluated. From each animal, approximately 5 mL of blood was collected by jugular venipuncture into a sterile tube without anticoagulant to obtain serum. Serum concentrations of total protein, albumin, creatinine, urea, total cholesterol, triglycerides, calcium, and phosphorus; and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, and gamma-glutamyl transferase (GGT) were determined on a automated multichannel analyzer. There were significant differences (p 0.05) in serum concentrations for total protein, globulin, albumin/globulin ratio (A:G), creatinine, urea, phosphorus, calcium/phosphorus ratio (Ca:P), AST, and GGT between Senepol calves and crossbred Holstein x Gir calves, but no significant differences in serum concentrations were observed between male and female calves for either breed (p > 0.05). It was concluded that the serum biochemistry profile of newborn Senepol and crossbred Holstein x Gir calves was affected by...

Objetivou-se avaliar as variações fisiológicas no perfil bioquímico sérico de bezerros neonatos da raça Senepol e mestiços Holandês x Gir de três a cinco dias idade, no município de Uberlândia, MG, e confrontar os valores entre as raças e sexos. Foram avaliados 168 bezerros neonatos, sendo 80 da raça Senepol (28 fêmeas e 52 machos) e 88 mestiços Holandês x Gir (49 fêmeas e 39 machos). Coletaram-se cinco mililitros de sangue, por venopunção da jugular externa, em tubos estéreis sem anticoagulante, para obtenção do soro. Foram determinadas as concentrações séricas de proteínas totais, albumina, creatinina, ureia, colesterol total, triglicérides, cálcio e fósforo; e as atividades de alanina aminotransferase (ALT), aspartato aminotransferase (AST), fosfatase alcalina e gama glutamiltransferase (GGT), em analisador automático multicanal. Confrontados os valores dos parâmetros avaliados entre a raça Senepol e os mestiços Holandês x Gir, as proteínas totais, globulinas, relação albumina e globulinas (A:G), creatinina, ureia, fósforo, relação cálcio e fósforo (Ca:P), AST e GGT apresentaram diferença (p0,05). Conclui-se que a diferença no manejo de ingestão de colostro e fatores inerentes à raça influenciam no perfil bioquímico sérico de bezerros neonatos e não há diferença nos valores dos parâmetros estudados para machos e fêmeas.

Serum biochemistry profile in newborn Senepol and crossbred Holstein x Gir calves aged three to five days in Uberlândia, Minas Gerais, Brazil / Perfil bioquímico sérico de bezerros neonatos da raça Senepol e mestiços Holandês x Gir de três a cinco dias de idade no município de Uberlândia, MG, Brasil

This study aimed to evaluate the physiological variations in the serum biochemical profile of neonate (3-5 days old) Senepol and crossbreed Holstein x Gir calves in Uberlândia, state of Minas Gerais, Brazil, and to compare values between breeds and sexes. In total, 168 healthy Senepol (n = 80, 28 females and 52 males) and crossbred Holstein x Gir (n = 88, 49 females and 39 males) calves were evaluated. From each animal, approximately 5 mL of blood was collected by jugular venipuncture into a sterile tube without anticoagulant to obtain serum. Serum concentrations of total protein, albumin, creatinine, urea, total cholesterol, triglycerides, calcium, and phosphorus; and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, and gamma-glutamyl transferase (GGT) were determined on a automated multichannel analyzer. There were significant differences (p 0.05) in serum concentrations for total protein, globulin, albumin/globulin ratio (A:G), creatinine, urea, phosphorus, calcium/phosphorus ratio (Ca:P), AST, and GGT between Senepol calves and crossbred Holstein x Gir calves, but no significant differences in serum concentrations were observed between male and female calves for either breed (p > 0.05). It was concluded that the serum biochemistry profile of newborn Senepol and crossbred Holstein x Gir calves was affected by...(AU)

Objetivou-se avaliar as variações fisiológicas no perfil bioquímico sérico de bezerros neonatos da raça Senepol e mestiços Holandês x Gir de três a cinco dias idade, no município de Uberlândia, MG, e confrontar os valores entre as raças e sexos. Foram avaliados 168 bezerros neonatos, sendo 80 da raça Senepol (28 fêmeas e 52 machos) e 88 mestiços Holandês x Gir (49 fêmeas e 39 machos). Coletaram-se cinco mililitros de sangue, por venopunção da jugular externa, em tubos estéreis sem anticoagulante, para obtenção do soro. Foram determinadas as concentrações séricas de proteínas totais, albumina, creatinina, ureia, colesterol total, triglicérides, cálcio e fósforo; e as atividades de alanina aminotransferase (ALT), aspartato aminotransferase (AST), fosfatase alcalina e gama glutamiltransferase (GGT), em analisador automático multicanal. Confrontados os valores dos parâmetros avaliados entre a raça Senepol e os mestiços Holandês x Gir, as proteínas totais, globulinas, relação albumina e globulinas (A:G), creatinina, ureia, fósforo, relação cálcio e fósforo (Ca:P), AST e GGT apresentaram diferença (p0,05). Conclui-se que a diferença no manejo de ingestão de colostro e fatores inerentes à raça influenciam no perfil bioquímico sérico de bezerros neonatos e não há diferença nos valores dos parâmetros estudados para machos e fêmeas.(AU)

Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro.

Current studies find that degenerated cartilage endplates (CEP) of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA) was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.

Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro

Current studies find that degenerated cartilage endplates (CEP) of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA) was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.

Testing genotoxicity and cytotoxicity strategies for the evaluation of commercial radiosterilized fetal calf sera.

Effects of 18 commercial lots of fetal calf serum (FCS) after gamma-irradiation and their non-irradiated counterparts were comparatively analyzed on CHO-K1 and MDBK MDL1 cells for genotoxicity [sister chromatid exchange (SCE), micronuclei (MNi), and single cell gel electrophoresis (SCGE)], cytotoxicity [cell-cycle progression (CCP), proliferative replication index (PRI), mitotic index (MI), growth promotion (GP), and plating efficiency (PE)], and microbiological properties (mycoplasma and bovine viral diarrhea virus contamination). SCE and SCGE were the most informative end-points for genotoxicity since significant differences were found in 44.4% (P<0.05-0.001, Student's t-test) and 61.1% (P<0.05-0.001, chi(2) test) samples, respectively. MI was the cytotoxicity assay revealing the greatest variation, showing differences in 66.7% (P<0.05-0.001, chi(2) test) samples. Thus, these three end-points for screening bioproducts such as FCS were found most suitable for detecting potential geno-cytotoxicants in biological samples; their simultaneous use could be strongly recommended.

The role of p38 mitogen-activated protein kinase in serum-induced leukemia inhibitory factor secretion by bone marrow stromal cells from pediatric myelodysplastic syndromes.

Stromal cells from pediatric myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) associated with MDS (MDS-AML) present high expression of leukemia inhibitor factor (LIF). We demonstrated using mitogen-activated protein kinase (MAPK) inhibitors that in stromal cells from pediatric MDS and MDS-AML, p38MAPK was critical in serum-induced secretion of LIF. The serum induction of phosphorylated p38MAPK form was observed only in stromal cells from healthy children, whereas in MDS and MDS-AML basal levels were maintained suggesting constitutive p38MAPK activation. Our study suggested the possible importance in pediatric MDS of p38MAPK signaling pathway which may be a future therapeutic target.

Mitochondrial calcium overload triggers complement-dependent superoxide-mediated programmed cell death in Trypanosoma cruzi.

The epimastigote stage of Trypanosoma cruzi undergoes PCD (programmed cell death) when exposed to FHS (fresh human serum). Although it has been known for over 30 years that complement is responsible for FHS-induced death, the link between complement activation and triggering of PCD has not been established. We have previously shown that the mitochondrion participates in the orchestration of PCD in this model. Several changes in mitochondrial function were described, and in particular it was shown that mitochondrion-derived O(2)(*-) (superoxide radical) is necessary for PCD. In the present study, we establish mitochondrial Ca(2+) overload as the link between complement deposition and the observed changes in mitochondrial physiology and the triggering of PCD. We show that complement activation ends with the assembly of the MAC (membrane attack complex), which allows influx of Ca(2+) and release of respiratory substrates to the medium. Direct consequences of these events are accumulation of Ca(2+) in the mitochondrion and decrease in cell respiration. Mitochondrial Ca(2+) causes partial dissipation of the inner membrane potential and consequent mitochondrial uncoupling. Moreover, we provide evidence that mitochondrial Ca(2+) overload is responsible for the increased O(2)(*-) production, and that if cytosolic Ca(2+) rise is not accompanied by the accumulation of the cation in the mitochondrion and consequent production of O(2)(*-), epimastigotes die by necrosis instead of PCD. Thus our results suggest a model in which MAC assembly on the parasite surface allows Ca(2+) entry and its accumulation in the mitochondrion, leading to O(2)(*-) production, which in turn constitutes a PCD signal.

Característica histológica, ultra-estrutural e produção de nitrito de folículos pré-antrais caprinos cultivados in vitro na ausência ou presença de soro / Histological and ultrastructural feature and nitrite production of caprine preantral follicles in vitro cultured in the presence or absence of serum

Avaliou-se o efeito da adição de diferentes tipos e concentrações de soro sobre o desenvolvimento e a sobrevivência de folículos ovarianos pré-antrais (FOPA) caprinos in vitro. Além disso, verificou-se a relação entre as concentrações de nitrito presentes no meio de cultivo e a viabilidade folicular. Cada par ovariano foi dividido em 29 fragmentos, sendo um destinado ao controle. Os fragmentos foram cultivados por um ou sete dias em meio essencial mínimo suplementado (MEM+) ou MEM+ com diferentes concentrações (10 ou 20 por cento) de soro fetal bovino (SFB), soro de cabra em estro (SCE) ou soro de cabra em diestro (SCD). Na análise morfológica após sete dias, apenas o tratamento com 10 por cento de SFB apresentou percentual de FOPA normais similar ao MEM+ (P>0,05). A análise ultra-estrutural dos folículos cultivados por sete dias com MEM+ ou MEM+ com 10 por cento de SFB mostrou danos oocitários, porém células da granulosa normais. A análise do meio de cultivo revelou correlação positiva entre a viabilidade folicular e a produção de nitrito. A suplementação com soro não melhorou a viabilidade de FOPA e a concentração de nitrito no meio de cultivo funcionou como um indicador da viabilidade das células da granulosa de FOPA caprinos cultivados in vitro.(AU)

The effect of the addition of different types and concentrations of sera on the viability and development of caprine preantal follicles (PAF) in vitro cultured was analyzed. In addition, it was evaluated the correlation between nitrite concentrations in culture medium and folicular viability. Each ovarian pair was divided in 29 fragments and one was used as control. The fragments were cultured for one or seven days in minimal essential medium (MEM+) or MEM+ with different concentrations of (10 or 20 percent) bovine fetal serum (BFS), estrous goat serum (EGS), or diestrous goat serum (DGS). After seven days, the morphological analysis showed that only the treatment with 10 percent BFS maintained the percentage of normal PAF similar to MEM+ (P>0.05). The ultrastructural analysis of follicles cultured for seven days in MEM+ or MEM+ with 10 percent BFS showed some oocyte damage, although the granulosa cells were normal. Analysis of culture medium revealed a positive correlation between follicular viability and nitrite production. Supplementation with serum did not improve the viability of PAF and nitrite levels in culture medium served as an indicator of viability of granulose cells from caprine PAF in vitro cultured.(AU)

Característica histológica, ultra-estrutural e produção de nitrito de folículos pré-antrais caprinos cultivados in vitro na ausência ou presença de soro / Histological and ultrastructural feature and nitrite production of caprine preantral follicles in vitro cultured in the presence or absence of serum

Avaliou-se o efeito da adição de diferentes tipos e concentrações de soro sobre o desenvolvimento e a sobrevivência de folículos ovarianos pré-antrais (FOPA) caprinos in vitro. Além disso, verificou-se a relação entre as concentrações de nitrito presentes no meio de cultivo e a viabilidade folicular. Cada par ovariano foi dividido em 29 fragmentos, sendo um destinado ao controle. Os fragmentos foram cultivados por um ou sete dias em meio essencial mínimo suplementado (MEM+) ou MEM+ com diferentes concentrações (10 ou 20 por cento) de soro fetal bovino (SFB), soro de cabra em estro (SCE) ou soro de cabra em diestro (SCD). Na análise morfológica após sete dias, apenas o tratamento com 10 por cento de SFB apresentou percentual de FOPA normais similar ao MEM+ (P>0,05). A análise ultra-estrutural dos folículos cultivados por sete dias com MEM+ ou MEM+ com 10 por cento de SFB mostrou danos oocitários, porém células da granulosa normais. A análise do meio de cultivo revelou correlação positiva entre a viabilidade folicular e a produção de nitrito. A suplementação com soro não melhorou a viabilidade de FOPA e a concentração de nitrito no meio de cultivo funcionou como um indicador da viabilidade das células da granulosa de FOPA caprinos cultivados in vitro.

The effect of the addition of different types and concentrations of sera on the viability and development of caprine preantal follicles (PAF) in vitro cultured was analyzed. In addition, it was evaluated the correlation between nitrite concentrations in culture medium and folicular viability. Each ovarian pair was divided in 29 fragments and one was used as control. The fragments were cultured for one or seven days in minimal essential medium (MEM+) or MEM+ with different concentrations of (10 or 20 percent) bovine fetal serum (BFS), estrous goat serum (EGS), or diestrous goat serum (DGS). After seven days, the morphological analysis showed that only the treatment with 10 percent BFS maintained the percentage of normal PAF similar to MEM+ (P>0.05). The ultrastructural analysis of follicles cultured for seven days in MEM+ or MEM+ with 10 percent BFS showed some oocyte damage, although the granulosa cells were normal. Analysis of culture medium revealed a positive correlation between follicular viability and nitrite production. Supplementation with serum did not improve the viability of PAF and nitrite levels in culture medium served as an indicator of viability of granulose cells from caprine PAF in vitro cultured.

Manutenção in vitro de células IDE8 em dois tipos de soro bovino / In vitro maintenance of IDE8 cells using two types of bovine serum

The present study had the objective of defining the culture conditions, optimizing the maintenance and expansion of an IDE-8 cell line in Brazil, with the aim to propose its use as a model for in vitro infection and multiplication of Brazilian strains of rickettsia and other hemoparasites. The supplementation of IDE-8 cells with two distinct fetal bovine sera (a Brazilian and an imported) was evaluated. Culture media were changed weekly and subcultures were carried out every 15 days. The development of cultures and subcultures was evaluated by the percentage of viability and cellular morphology. The results indicate that the imported SFB can be replaced by the Brazilian SFB one, as no significant differences (P<0.05) were seen among culture viabilities.(AU)

Serum biochemistry of an, atlantic Yellow-Nosed Albatross Thalassarche chlororhynchos (Gmelin, 1789) / Bioquímica do soro sangüíneo de um albatroz Thalassarche chlororhynchos (Gmelin, 1789)

Colheu-se uma amostra de sangue de um albatroz Thalassarche chlororhynchos (Gmelin, 1789), recolhido pela Polícia do Meio Ambiente de Ubatuba, SP, para análises da bioquímica do soro. O objetivo dessas análises foi o de apontar possíveis alterações dos parâmetros bioquímicos, visando facilitar futuras casas para reabilitação de aves. Nenhuma anormalidade foi encontrada e o animal morreu após três dias da captura.(AU)

Serum biochemistry of an, atlantic Yellow-Nosed Albatross Thalassarche chlororhynchos (Gmelin, 1789)

Colheu-se uma amostra de sangue de um albatroz Thalassarche chlororhynchos (Gmelin, 1789), recolhido pela Polícia do Meio Ambiente de Ubatuba, SP, para análises da bioquímica do soro. O objetivo dessas análises foi o de apontar possíveis alterações dos parâmetros bioquímicos, visando facilitar futuras casas para reabilitação de aves. Nenhuma anormalidade foi encontrada e o animal morreu após três dias da captura.

Manutenção in vitro de células IDE8 em dois tipos de soro bovino / In vitro maintenance of IDE8 cells using two types of bovine serum

The present study had the objective of defining the culture conditions, optimizing the maintenance and expansion of an IDE-8 cell line in Brazil, with the aim to propose its use as a model for in vitro infection and multiplication of Brazilian strains of rickettsia and other hemoparasites. The supplementation of IDE-8 cells with two distinct fetal bovine sera (a Brazilian and an imported) was evaluated. Culture media were changed weekly and subcultures were carried out every 15 days. The development of cultures and subcultures was evaluated by the percentage of viability and cellular morphology. The results indicate that the imported SFB can be replaced by the Brazilian SFB one, as no significant differences (P<0.05) were seen among culture viabilities.

Increased NHE1 expression is associated with serum deprivation-induced differentiation in immortalized rat proximal tubule cells.

We studied the proton secretion mechanisms involved with pHi regulation in immortalized rat proximal tubule cells (IRPTC), a SV40-immortalized cell line derived from rat proximal tubule, and characterized the effects of serum deprivation on them. Using pHi measurements with the fluorescent probe BCECF, we demonstrated that the IRPTC express both Na+/H+ exchanger and H+-ATPase, but only NHE1 is modulated by serum deprivation. In these cells, 24 h of serum starvation increased pHi from 7.08+/-0.008 (n=34) to 7.18+/-0.018 (n=33) as well as the pH recovery rate from intracellular acidification with NH4Cl from 0.29+/-0.022 pH U/min (n=14) to 0.50+/-0.024 pH U/min (n=14), without modifying their buffering capacity. These effects were followed by several modifications in morphological features, indicating an increase in differentiation status. The altered activity of NHE1 was consistent with an increase of both transcription and translation of the antiporter, as the utilization of actinomycin D and cycloheximide significantly inhibited the upregulation of NHE1 induced by serum withdrawal. Inhibition of tyrosine phosphorylation by genistein blocked the serum deprivation-dependent activation of NHE. Moreover, the pharmacological inhibition of MEK1/2, the upstream activator of ERK1/2 by UO-126, significantly inhibited the stimulatory effect of serum starvation on Na+/H+ exchanger activity, whereas the putative p38 MAPK inhibitor SB-203580 failed to cause any effect on pHi recovery rates. Our findings indicate that during IRPTC differentiation by serum deprivation, there was a net enhancement of NHE1 activity. This upregulation of NHE by serum removal was consistent with an increase of RNA and protein synthesis of the exchanger, which depends on tyrosine kinase phosphorylation and ERK pathway activation.

A randomized controlled trial of fluid supplementation in term neonates with severe hyperbilirubinemia.

OBJECTIVE: To evaluate the effectiveness of fluid supplementation in decreasing the rate of exchange transfusion and the duration of phototherapy in term neonates with severe nonhemolytic hyperbilirubinemia. STUDY DESIGN: This was a randomized controlled trial conducted in a tertiary care referral unit in northern India. Seventy-four term neonates with severe nonhemolytic hyperbilirubinemia (total serum bilirubin > 18 mg/dL [308 micromol/L] to < 25 mg/dL [427 micromol/L]). The subjects were randomized to an "extra fluids" group (intravenous fluid supplementation for 8 hours and oral supplementation for the duration of phototherapy; n = 37) or a control group (n = 37). RESULTS: At inclusion, 54 infants (73%) had high serum osmolality, including 28 (75%) in the extra fluids group and 26 (70%) in the control group. The proportion of infants who underwent exchange transfusion was lower in the extra fluids group than in the control group: 6 (16%) versus 20 (54%)(P = .001; relative risk = 0.30; 95% confidence interval = 0.14 to 0.66). The duration of phototherapy was also shorter in the extra fluids group: 52 +/- 18 hours versus 73 +/- 31 hours (P = .004). CONCLUSION: Fluid supplementation in term neonates presenting with severe hyperbilirubinemia decreased the rate of exchange transfusion and duration of phototherapy.

Dexamethasone and fetal calf serum effects in differentiation of Vero cells cultured on type I collagen gel.

Vero cells, a fibroblastic cell line, were cultured on a collagen I gel without fetal calf serum (FCS), with the addition of 10% FCS, 10% FCS plus dexamethasone (DEX) or 20% FCS. Our objective was to determine the effects of DEX on the differentiation pattern of fibroblastic cells cultured on a collagen substrate. We found that cells cultured with or without 10% FCS were capable of migrating into the collagen matrix. Near the cells in the gel, we found the deposition of extracellular granulations. Cytochemical data suggests that this material is glycosaminoglycans and/or proteoglycans. Surrounding the cells, a fibronectin deposition was found in the collagen. Thus, these cells make up a structure similar to a loose connective tissue. On the other hand, cells cultured with 10% FCS plus DEX or with 20% FCS did not invade the collagen matrix but formed multiple cell layers poor in fibronectin. On collagen I gel surface, we found an acellular layer rich in collagen IV, which appeared between the cells and the substrate. Thus, DEX or 20% FCS, furnished to the cells cultured on a collagen gel, block cell migration into the substrate and induce them to produce a basement membrane-like structure.